Photosynthesis Research

, Volume 84, Issue 1–3, pp 145–151 | Cite as

Quantification of non-Q B -reducing centers in leaves using a far-red pre-illumination

  • Gert SchanskerEmail author
  • Reto J. Strasser
Regular paper


An alternative approach to quantification of the contribution of non-Q B -reducing centers to Chl a fluorescence induction curve is proposed. The experimental protocol consists of a far-red pre-illumination followed by a strong red pulse to determine the fluorescence rise kinetics. The far-red pre-illumination induces an increase in the initial fluorescence level (F25 μs) that saturates at low light intensities indicating that no light intensity-dependent accumulation of Q A occurs. Far-red light-dose response curves for the F25 μs-increase give no indication of superimposed period-4 oscillations. F25 μs-dark-adaptation kinetics following a far-red pre-pulse, reveal two components: a faster one with a half-time of a few seconds and a slower component with a half-time of around 100 s. The faster phase is due to the non-Q B -reducing centers that re-open by recombination between Q A and the S-states on the donor side. The slower phase is due to the recombination between Q B and the donor side in active PS II reaction centers. The pre-illumination-induced increase of the F25 μs-level represents about 4–5% of the variable fluorescence for pea leaves (∼2.5% equilibrium effect and 1.8–3.0% non-Q B -reducing centers). For the other plant species tested these values were very similar. The implications of these values will be discussed.


far-red light non-QB-reducing centers OJIP-transient plant species dependence QA–QB equilibrium effect 






F25 μ s

initial fluorescence, for dark adapted leaves equal to Fo


intermediate fluorescence induction level when measured at low light intensity


variable fluorescence here used as maximum variable fluorescence


light emitting diode

OJIP curve

fluorescence induction transient defined by the names of its intermediate steps, O-level is fluorescence-level at 25 μs, J-level is fluorescence-level at ∼2 ms, I-level is fluorescence level at ∼30 ms and P-level is F m , the maximum fluorescence

QA and QB

primary and secondary quinone electron acceptors of Photosystem II


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  1. Black, MT, Brearley, T, Horton, P 1986Heterogeneity in chloroplast photosystem IIPhotosynth Res8193207CrossRefGoogle Scholar
  2. Chylla, RA, Whitmarsh, J 1989Inactive photosystem II complexes in leavesPlant Physiol90765772Google Scholar
  3. Crofts, AR, Robinson, HH, Snozzi, M 1984Reactions of quinones at catalytic sites; a diffusional role in H-transferSybesma, C eds. Advances in Photosynthesis Research, Vol I.Martinus Nijhoff/Dr W Junk PublishersThe Hague, The Netherlands461468Google Scholar
  4. Dekker, JP, Gorkom, HJ, Wensink, J, Ouwehand, L 1984Absorbance difference spectra of the successive redox states of the oxygen-evolving apparatus of photosynthesisBiochim Biophys Acta76719Google Scholar
  5. Diner, BA 1977Dependence of the deactivation reactions of Photosystem II on the redox state of plastoquinone pool A varied under anaerobic conditions; equilibria on the acceptor side of Photosystem IIBiochim Biophys Acta460247258PubMedGoogle Scholar
  6. Diner, BA, Petrouleas, V, Wendoloski, JJ 1991The iron-quinone electron acceptor complex of Photosystem IIPhysiol Plant81423436CrossRefGoogle Scholar
  7. Forbush, B, Kok, B 1968Reaction between primary and secondary electron acceptors of Photosystem II of photosynthesisBiochim Biophys Acta162243253PubMedGoogle Scholar
  8. Guenther, JE, Melis, A 1990The physiological significance of Photosystem II heterogeneity in chloroplastsPhotosynth Res23105109CrossRefGoogle Scholar
  9. Hauser, M, Eichelmann, H, Oja, V, Heber, U, Laisk, A 1995Stimulation by light of rapid pH regulation in the chloroplast stroma in vivo as indicated by CO2 solubilization in leavesPlant Physiol10810591066PubMedGoogle Scholar
  10. Hsu, B-D, Lee, J-Y 1991Characterization of the Photosystem II centers inactive in plastoquinone reduction by fluorescence inductionPhotosynth Res27143150CrossRefGoogle Scholar
  11. Hsu, B-D, Lee, J-Y, Jang, Y-R 1989A method for analysis of fluorescence induction curve from DCMU-poisoned chloroplastsBiochim Biophys Acta9754449Google Scholar
  12. Lavergne, J 1991Improved UV-visible spectra of the S-transitions in the photosynthetic oxygen-evolving systemBiochim Biophys Acta1060175188Google Scholar
  13. Lavergne, J, Briantais, J-M 1996Photosystem II heterogeneityOrt, DRYocum, CF eds. Oxygenic Photosynthesis: The Light Reactions, Advances in Photosynthesis, Vol 4.Kluwer Academic PublishersDordrecht/Boston/London265287Google Scholar
  14. Lavergne, J, Etienne, A-L 1980Prompt and delayed fluorescence of chloroplasts upon mixing with dichlorophenylmethylureaBiochim Biophys Acta593136148PubMedGoogle Scholar
  15. Lavergne, J, Leci, E 1993Properties of inactive Photosystem II centersPhotosynth Res35323343CrossRefGoogle Scholar
  16. Melis, A 1985Functional properties of Photosystem IIβ in spinach chloroplastsBiochim Biophys Acta808334342Google Scholar
  17. Renger, G, Hagemann, R, Fromme, R 1986The susceptibility of the p-benzoquinone-mediated electron transport and atrazin binding to trypsin and its modification by CaCl2 in thylakoids and PS II membrane fragmentsFEBS Lett203210214CrossRefGoogle Scholar
  18. Renger, G, Fromme, R, Hagemann, R 1988The modification of atrazine binding by the redox state of the endogenous high-spin iron and by specific proteolytic enzymes in Photosystem II membrane fragments and intact thylakoidsBiochim Biophys Acta935173183Google Scholar
  19. Robinson, HH, Crofts, AR 1984Kinetics of proton uptake and the oxidation-reduction reactions of the quinone acceptor complex of PS II from pea chloroplastsSybesma, C eds. Advances in Photosynthesis Research, Vol I.Martinus Nijhoff/Dr W. Junk PublishersThe Hague, The Netherlands477480Google Scholar
  20. Rutherford, AW, Inoue, Y 1984Oscillation of delayed luminescence from PS II: recombination of S2Q B and S3Q B FEBS Lett165163170CrossRefGoogle Scholar
  21. Rutherford, AW, Govindjee, , Inoue, Y 1984Charge accumulation and photochemistry in leaves studied by thermoluminescence and delayed light emissionProc Natl Acad Sci USA8111071111Google Scholar
  22. Schansker, G, Goussias, C, Petrouleas, V, Rutherford, AW 2002Reduction of the Mn cluster of the water-oxidizing enzyme by nitric oxide: formation of an S−2 stateBiochemistry4130573064PubMedGoogle Scholar
  23. Strasser, RJ, Tsimilli-Michael, M, Srivastava, A 2005Analysis of the chlorophyll a transientPapageorgiou, GGovindjee,  eds. Chlorophyll Fluorescence: A Signature of Photosynthesis.Kluwer Academic PublishersDordrecht/Boston/London321362Google Scholar
  24. Tomek, P, Ilik, P, Lazar, D, Stroch, M, Naus, J 2003On the determination of Q B -non-reducing photosystem II centers from chlorophyll a fluorescence inductionPlant Sci164665670CrossRefGoogle Scholar
  25. Vermaas, WFJ, Renger, G, Dohnt, G 1984The reduction of the oxygen-evolving system in chloroplasts by thylakoid componentsBiochim Biophys Acta764194202Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Bioenergetics LaboratoryUniversity of GenevaGenevaSwitzerland

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